• Title/Summary/Keyword: Phosphate stress

Search Result 199, Processing Time 0.03 seconds

Effect of Promoters on the Heme Production in a Recombinant Corynebacterium glutamicum (재조합 Corynebacterium glutamicum으로부터 헴첼 생산에 미치는 프로모터의 효과)

  • Yang, Hyungmo;Kim, Pil
    • Microbiology and Biotechnology Letters
    • /
    • v.47 no.3
    • /
    • pp.337-342
    • /
    • 2019
  • We published that bacterial heme was over-produced in a recombinant Corynebacterium glutamicum expressing 5-aminolevulinic acid synthase ($hemA^+$) under control of a constitutive promoter ($P_{180}$) and the heme-producing C. glutamicum had commercial potentials; as an iron feed additive for swine and as a preservative for lactic acid bacteria. To enhance the heme production, the $hemA^+$ gene was expressed under controls of various promoters in the recombinant C. glutamicum. The $hemA^+$ expression by $P_{gapA}$ (a constitutive glycolytic promoter of glyceraldehyde-3-phosphate dehydrogenase) led 75% increase of heme production while the expression by $P_{H36}$ (a constitutive, very strong synthetic promoter) resulted in 50% decrease compared with the control ($hemA^+$ expression by $P_{180}$ constitutive promoter). The $hemA^+$ expression by a late log-phase activating $P_{sod}$ (an oxidative-stress responding promoter of superoxide dismutase) led 50% greater heme production than the control. The $hemA^+$ expression led by a heat-shock responding chaperone promoter ($P_{dnaK}$) resulted in 121% increase of heme production at the optimized heat-shock conditions. The promoter strength and induction phase are discussed based on the results for the heme production at an industrial scale.

Seasons affect the phosphorylation of pork sarcoplasmic proteins related to meat quality

  • Zeng, Xianming;Li, Xiao;Li, Chunbao
    • Animal Bioscience
    • /
    • v.35 no.1
    • /
    • pp.96-104
    • /
    • 2022
  • Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P2b, P21, and P22) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P21) and positive association with free water (P22). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

Qualitative and quantitative assessment of process related impurities in Brigatinib raw material and formulations using HPLC

  • Attada Tharun;Potnuru Jagadeesh;B Srinivasa Kumar;Kota Thirumala Prasad;Venkateswara Rao Anna
    • Analytical Science and Technology
    • /
    • v.36 no.4
    • /
    • pp.180-190
    • /
    • 2023
  • The presence of process related impurities in any drug or the drug product was associated with its safety, stability and efficacy. The overall literature survey proved that there is no method published on the assessment of process related impurities in brigatinib. In this study, a simple, reliable and stable HPLC qualitative method was reported for quantification of process related impurities with easy and quick extraction procedure. The impurities along with standard brigatinib was resolved on Lichrospher® C18 (250 mm × 4.6 mm; 5 ㎛ particle size) column in room temperature using methanol, acetonitrile, pH 4.5 phosphate buffer in 55:25:20 (v/v) at 1.0 mL/min as mobile phase and UV detection at 261 nm. The method produces well resolved peaks at retention time of 4.60 min, 12.28 min, 3.37 min, 7.34 min and 8.39 min respectively for brigatinib, impurity A, B, C and D. The method produces a very sensitive detection limit of 0.0065 ㎍/mL, 0.0068 ㎍/mL, 0.0053 ㎍/mL and 0.0058 ㎍/mL for impurity A, B, C and D respectively with calibration curve linear in the concentration range of 22.5-135 ㎍/mL for brigatinib and 0.0225-0.135 ㎍/mL for impurities. The method produces all the validation parameters under the acceptable level and doesn't produces any considerable changes in peak area response while minor changes in the developed method conditions. The method can effectively resolve the unknown stress degradation products along with known impurities with less % degradation. The method can efficiently resolve and quantify the impurities in formulation and hence can suitable for the routine quality analysis of brigatinib in raw material and formulation.

Novel stability indicating high-performance liquid chromatography method for the separation and simultaneous quantification of acalabrutinib and its impurities in pharmaceutical formulation

  • Venu Gopal Kamani;Sujatha M;Guna Bhushana Daddala
    • Analytical Science and Technology
    • /
    • v.36 no.1
    • /
    • pp.32-43
    • /
    • 2023
  • This study reports for the first time about a stability indicating RP-HPLC method for qualitative and quantitative determination of acalabrutinib in bulk and dosage form and in presence its impurities 1, 2 and 3. The chromatographic separation was carried on Zorbax XDB-C18 (250×4.6 mm; 5 µ id) as stationary phase, Phosphate buffer pH 6.4 and methanol 80:20 (v/v) as mobile phase at a flow rate of 1.0 mL/min, UV detection was carried at wavelength of 238 nm and the analysis was completed with a run time of 15 min. In these conditions the retention time of acalabrutinib and its impurities 1, 2 and 3 was observed to be 3.50, 4.83, 8.40 and 9.93 min respectively. The method was validated for system suitability, range of analysis, precision, specificity, stability and robustness. Spiked recovery at 50 %, 100 % and 150 % was carried for both standard and impurities and the acceptable % recovery of 98-102 was observed for acalabrutinib and both impurities studied and the % RSD in each spiked level was found to be less than 2. Stability tests were done through exposure of the analyte solution to five different stress conditions i.e expose to 1N hydrochloric acid, 1 N sodium hydroxide, 3 % peroxide, 80 ℃ temperature and UV radiation at 254 nm. In all the degradation condition, standard drug acalabrutinib was detected along with both the impurities studied and the degradation products were successfully separated. In the formulation analysis there is no other chromatographic detection of other impurities and formulation excipients. Hence the developed method was found to be suitable for the quantification of acalabrutinib and can separate and analyse impurities 1 and 2.

Effects of Phenolic and Phosphite Antioxidants on the properties for PC/ABS Blends during High-Shear-Rate Processing (고속 전단 가공에서 페놀계와 인산계 산화방지제에 의한 PC/ABS 블렌드의 물성 변화 연구)

  • Lee, Han Ki;Kim, Seon Hong;Lee, Hyung Il;Yoo, Jae Jung;Yong, Da Kyoung;Choi, Seok Jin;Lee, Seung Goo;Lee, Kee Yoon
    • Korean Chemical Engineering Research
    • /
    • v.52 no.2
    • /
    • pp.266-271
    • /
    • 2014
  • The effects of antioxidants on the properties of Polycarbonate/Acrylonitrile-Butadiene-Styrene(PC/ABS) blends were studied for the functions of the screw speed and loaded duration of high shear rate processing in order to investigate the degradation for PC/ABS blends. Tris-(2,4-di-tert-butyl-phenyl phosphate) (A1) and Bis(2,4-dicumylphenyl) pentaerythritol diphosphite (A3) as phosphite antioxidants and Octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propionate (A2) as a phenolic antioxidant are used. The thermal properties were detected by TGA and severely decreased, after the processing. The stress-induced and thermal degradation for PC/ABS blends with the antioxidant A3 was retarded better than the others. By using UTM, the mechanical properties also showed individually decreased according to the antioxidants, after the processing, especially, the elongations showed considerable decline behaviors, while the tensile strengths of PC/ABS blends changed very little. For example, in the operating conditions of 1000rpm of screw speed and 20 seconds of loaded period, the elongations decreased from 148% before the processing, to 91.6% with the A1, to 63% with the A2 and to 131% with the A3 after the processing, respectively. In order to get the morphological properties, the size distributions of the dispersed phases for PC/ABS were investigated by SEM analysis and tended to decrease, as the screw speed and loaded period of the processing increased. Therefore, we confirmed that the antioxidant A3 was the best of all of three to inhibit the stress-induced degradation of PC/ABS blends during the high shear rate processing.

A Comparative Study on the Mechanical Properties of Plywood treated with Several Fire Retardant Chemicals (I) - Effect of Soaking Time on the Static Bending Strength of Treated Plywood - (수종(數種) 내화약제(耐火藥劑)로 처리(處理)된 합판(處理)의 기술적(技術的) 성질(性質)에 관(關)한 비교연구(比較硏究)(I) - 처리합판(處理處理)의 휨강도(强度)에 미치는 침지시간(浸漬時間)의 영향(影響) -)

  • Kim, Jong-Man;Chung, Woo-Yang;Lee, Phil-Woo
    • Journal of the Korean Wood Science and Technology
    • /
    • v.12 no.2
    • /
    • pp.20-26
    • /
    • 1984
  • This study was carried out to investigate the influence of chemical type and its retention in the fire-retardant treated plywoods on the static bending strength, a property peculiar to plywood. Being soaked in 20% aqueous solution of $(NH_4)_2SO_4$, $NH_4H_2PO_4$, $(NH_4)_2HPO_4$. Borax-Boric acid and Minalith for 3 to 12 hours at three-hour intervals and redried at $120^{\circ}C$ in hot press, the treated plywoods were put to static bending test. The values of chemical treated plywoods in Stress at proportional limit, Modulus of elasticity, Modulus of rupture and Work per unit volume to proportional limit were widely higher than those of water treated plywoods(control) and Borax-Boric acid treatment showed the highest value in the four mechanical data. And the bending strength of fire-retardant treated plywoods increased with the extension of soaking time or the increase of chemical retention in themselves. Borix-Boric acid was the desirable fire-retardant for thin plywood in view of mechanical strength and soaking defects in this study.

  • PDF

Effects of Fermented Soybean Meal on Immune Response of Weaned Calves with Experimentally Induced Lipopolysaccharide Challenge

  • Kwon, In-Hyuk;Kim, Myung-Hoo;Yun, Cheol-Heui;Go, Jong-Yeol;Lee, Chan-Ho;Lee, Hyun-June;Phipek, Wisut;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.24 no.7
    • /
    • pp.957-964
    • /
    • 2011
  • The present study was conducted to evaluate the effects of fermented soybean meal (FSBM) on the level of cortisol hormone and immune-related serum proteins in weaned calves after experimentally induced lipopolysaccharide (LPS) challenge. Holstein neonatal calves (n = 21; 8 males and 13 females, BW = $42.2{\pm}6.15$ kg) were randomly allocated to one of two dietary treatments: SBM (control calf starter having soybean meal (SBM) as a main protein source) and FSBM (substitute SBM in control diet with FSBM) groups. All calves were fed milk replacer using an automatic milk-feeder according to step-down milking method and weaned at 7 weeks old. Experimental diets were given to calves ad libitum throughout the experimental period. For LPS challenge, all calves except negative control animals given phosphate buffered saline (PBS), were injected subcutaneously with Salmonella typhimurium LPS on day 7 (D7) after weaning (D0). No significant difference in growth performance and milk intake was observed between SBM and FSBM calves. Feeding FSBM diet resulted in significantly (p<0.05) higher LPS-specific IgG at D12 and D19 and LPS-specific IgA at D19 in peripheral blood. Calves fed with FSBM diet also had significantly (p<0.05) higher concentration of serum haptoglobin (Hp) at D8. Overall concentration of cortisol in FSBM group was considerably lower than that of SBM group. Results from current study indicate that FSBM may provide beneficial effects in alleviating weaning stress and enhance immune status of weaned calves.

Effects of Acetamide and Lactamide on the Viability of Frozen-thawed Mammalian Cells (포유류배양세포 동결보존에 있어 Lactamide의 효과)

  • Kim, Hyun;Cho, Young Moo;Ko, Yeoung-Gyu;Seong, Hwan-Hoo
    • Journal of Life Science
    • /
    • v.24 no.11
    • /
    • pp.1252-1257
    • /
    • 2014
  • While dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant agent in the cryopreservation of cultured mammalian cells, it has been reported to cause differentiation of some cell lines by DNA methylation and associated histone modifications. To avoid the side effects of DMSO in cryopreservation, other agents might be more appropriate for maintaining the stable differentiation of cultured cell phenotypes through cryopreservation. All cryoprotectants should be highly soluble in water and display low cell toxicity. Cryoprotective agents have been shown to be effective in animal sperm preservation, and eight types of amides were examined in the cryopreservation of cultured mouse endothelial cells. Among the amides examined, acetamide and lactamide were effective cryoprotectants for cultured mammalian cells. The most effective concentration of lactamide, 1.5 M, had an even lower cryoprotective ability than 1M DMSO. Because successful cryopreservation of cultured cells is hampered by osmotic stress, the adequate ionic concentration was determined by diluting phosphate-buffered saline (PBS) in the 1.5M lactamide solution. The most effective concentration was $0.4{\times}PBS$, which minimized osmotic stress during the cryopreservation of cultured cells. As the addition of high molecular weight materials in cryopreservation media improves the viability of cells, the effects of bovine serum albumin (BSA), hydroxyethyl-starch (HES), and dextran were examined. The best combination of lactamide-based media for cryopreservation was found to be 1.5 M lactamide in $0.4{\times}PBS$ with 1% BSA.

Polyamine Biosynthesis in Red Pepper and Chinese Cabbage by the Application of Liquid Pig Manure (돈분뇨 액비시용에 의한 고추 및 배추의 polyamine 생합성)

  • Hwang, Seon-Woong;Sung, Jwa-Kyung;Kang, Bo-Ku;Lee, Choon-Soo;Yun, Seung-Gil;Kim, Tae-Wan;Eom, Ki-Cheol
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.37 no.3
    • /
    • pp.171-176
    • /
    • 2004
  • To investigate the availability of nitrogen decomposed and released from liquid pig manure (LPM), this experiment was performed with red pepper (Capsicum annuum L., cv. Hanbando), and Chinese cabbage (Brassioa campestris L., cv. Konaenggiyeureumbaechoo) in 2001. Based on the total nitrogen of chemical fertilizer, both red pepper and chinese cabbage were treated with three and four applications of LPM, respectively. Yield of red fruits in the red pepper was increased by an enhancement of LPM application. However, that of chinese cabbage was enhanced with a reducing supply of LPM. Biosynthesis of polyamine in both crops such as red pepper and chinese cabbage was large in the early growth stage and was not increased by LPM application. The high biosynthesis of bound polyamine, monoamine and diamine, in the early growth stage was changed in an increase of conjugated polyamine and polyamine with a process of crop growth. Inorganic components in the leaf of red pepper by LPM application were equal or slightly lower than in chemical fertilizer, however, from the middle growth stage, contents of phosphate and potassium were increased. Those of chinese cabbage were slightly decreased from the early growth stage to the late. Considering this experiment, the thoughtless supply of LPM has not resulted in certain crop damages, and an application of LPM to increase a yield was different from crop species at some extent.

Effect of Soil Salinity on Nitrate Accumulation of Lettuce (토양 염류집적이 상추의 Nitrate 축적에 미치는 영향)

  • Jin, Sun-Jae;Cho, Hyun-Jong;Chung, Jong-Bae
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.37 no.2
    • /
    • pp.91-96
    • /
    • 2004
  • Accumulation of nitrate in edible crops is undesirable due to potential risks to human health. Since nitrate has a role in the osmotic regulation of plants, salt accumulation in soil is expected to stimulate nitrate accumulation in plants. Lettuce (Lactuca sativa L.) was grown in soils of different salinities, 9.69 and $4.49dS\;m^{-1}$, in a greenhouse, and the effect of soil salinity on nitrate accumulation in lettuce was investigated. Content of nitrate in the lettuce increased significantly as soil salinity increased under low light intensity and ample supply of nitrate in root media. Soluble sugar and oxalate contents in lettuce were also significantly higher in the soil of higher salinity. Phosphate, Cl, and $SO_4$ contents in lettuce were not significantly different in soils of different salinities. Among the cations, K content in lettuce was significantly higher in the soil of higher salinity, but Na, Ca, and Mg comtents were not much influenced. Comparing to the lettuce grown in low salinity soil, although the growth of lettuce was decreased by 9% in the soil of higher salinity, nitrate accumulation in the lettuce was increased by 18.6%. These results indicate that higher nitrate content in lettuce of higher salinity soil is a positive accumulation to adapt to the water stress condition. The nitrate accumulation of vegetables grown in plastic film houses is known to be due to the heavy fertilization and low light intensity, but salt accumulation in the soil, which can lower soil water potential, is expected to stimulate the nitrate accumulation further.